Search Results (69)

National Tuberculosis Reference Laboratories (NTRLs) are central to tuberculosis (TB) control programs. Between 2018 and 2024, the Republic of Congo, a country of 6 million inhabitants, achieved a transformative strengthening of its TB diagnostic system, coordinated by the NTRL. Strategic investments, supported mainly by international partners, enabled a substantial decentralization of services, expanding the diagnostic network from 38 to 113 diagnostic and testing centers and increasing GeneXpert sites from 3 to 31. The expansion of the diagnostic network and specimen referral system was associated with a reduced structural gap in diagnostic coverage by extending access to GeneXpert testing to a larger number of peripheral and previously underserved centers. Critically, the establishment of a BSL-3 laboratory and the deployment of advanced assays like Xpert MTB/XDR ended the reliance on overseas testing by introducing in-country capacity for multidrug-resistant and pre-extensively drug-resistant TB detection. These systemic improvements were associated with significant positive outcomes, including an annual molecular testing surging from 11,609 in 2022 to over 27,000 in 2024 and bacteriological confirmation rates rising from 34 to 73%. This comprehensive laboratory systems strengthening, which also facilitated cross-programmatic initiatives like HIV and Mpox testing integration, underscores how sustained investment in infrastructure, logistics, and quality management is fundamental to improving case detection, surveillance, and progress toward the WHO End TB Strategy milestones. Full article

Tuberculosis is still one of the leading infectious causes of morbidity and mortality worldwide. Rapid diagnosis is essential for effective treatment and control of tuberculosis transmission. In recent years, nucleic acid amplification tests (NAATs), such as GeneXpert MTB/RIF, BD MAX™, Xpert MTB/RIF-Ultra, have significantly improved tuberculosis diagnostics. However, they mainly require expensive and advanced equipment. The aim of our study was to assess the usefulness of the novel RITA MTBC assay in this diagnostic context. A total of 61 clinical specimens were tested using the RITA MTBC assay in comparison with established molecular diagnostic platforms (GeneXpert and BD MAX™), used as molecular reference assays. Culture and microscopy were performed as part of initial clinical assessment, but the comparative analysis focused on molecular assays to provide a relevant evaluation of diagnostic performance. Among 31 samples previously identified as positive for M. tuberculosis DNA, the assay correctly detected 30 (LOT HPA01/20230601) and 29 (LOT HPA01/20230602). Of 30 negative samples, 28 and 30 were confirmed negative for the respective lots. These results correspond to an average sensitivity of 95% and an average specificity of 97%. The kit demonstrated diagnostic performance that meets requirements for molecular testing in tuberculosis, with sensitivity and specificity comparable to established platforms, although further validation on larger sample sets is necessary. Nevertheless, its excellent specificity, rapid turnaround time, and operational simplicity, make it especially well-suited for decentralized or resource-limited settings. These findings underscore the potential of RITA MTBC as a valuable diagnostic tool in both routine clinical settings and in populations with limited access to healthcare. Full article

Rifampicin-resistant tuberculosis (RR-TB) remains a major threat to global TB control, primarily driven by mutations in the rpoB gene of Mycobacterium tuberculosis (Mtb). Most resistance-conferring mutations occur within the 81-base pair RIF resistance determining region (RRDR), particularly at codons S450L, H445Y/D, and D435V, which are strongly associated with high level resistance. However, increasing evidence of low-frequency and disputed variants both within and beyond the RRDR reveals a broader genetic spectrum that contributes to diagnostic uncertainty and variable phenotypic outcomes. This review summarizes current knowledge of high frequency, low frequency, and disputed rpoB mutations and their implications for molecular detection of RIF resistance. Structural analyses show that specific amino acid substitutions alter key hydrogen bonds or create steric hindrance in the RIF-binding pocket, leading to diverse resistance levels. Despite the success of molecular platforms such as Xpert MTB/RIF and line probe assays, their hotspot-based detection limits sensitivity to noncanonical variants. Lowering the minimum inhibitory concentration (MIC) breakpoint and integrating sequencing-based approaches, such as targeted and whole-genome sequencing, can enhance detection accuracy. A combined genomic and phenotypic framework will be essential to close existing diagnostic gaps and advance precision guided management of RIF-resistant and multidrug-resistant tuberculosis. Full article

Background/Objectives: Detection of carbapenemases (KPC, OXA-48, VIM, IMP, NDM) from blood cultures (BCs) by standard methods takes 48–72 h and includes BC seeding, susceptibility testing and carbapenemase detection. Automated qPCR panels provide results in 1 h but are very costly. We aim to evaluate a low-cost and rapid immunochromatographic (IC) test directly from positive BCs using the reference method as a comparator. Methods: Ninety-one positive BCs from real-world patients and sixty-four simulated BCs were included. BC broth was treated with SDS and washed before analysis with the K.N.I.V.O. carbapenemase detection IC test. Discordant results were confirmed through the NG Carba-5 IC test and GeneXpert Carba-R qPCR test. Results: The test detected 100% of the 87 carbapenemase-producing BCs tested (sensitivity: 100% [CI95%: 95.8–100%]). However, 13 BCs generated false positive bands for NDM and/or OXA-48 (specificity: 80.8% [CI95%: 69.5–89.4%). The positive and negative predictive values were 87.0% (CI95%: 80.4–91.6%) and 100% (CI95%: 93.5–100%). Analysis of BCs providing false positive results through both confirmatory tests showed that BCs were negative for these carbapenemases. Conclusions: This is the first evaluation of the K.N.I.V.O. IC test directly from positive BCs, with a pragmatic confirmation algorithm using a second IC test or qPCR in case of NDM or OXA-48, that addresses K.N.I.V.O.’s specificity gap. The main limitation of this work is that confirmatory testing was performed only in false positives. The implementation of the K.N.I.V.O. IC test would contribute to early carbapenemase detection in BCs and is an alternative for low-resource hospitals where qPCR panels are not available. Full article

Background/Objectives: Multidrug-resistant (MDR) nosocomial infections remain a major challenge in intensive care units (ICUs), where delays in diagnosis and suboptimal antimicrobial therapy significantly impact outcomes. This narrative review synthesizes international literature and local epidemiological data from Western Romania to examine the role of rapid molecular diagnostics in the management of MDR infections and their integration with prevention and antimicrobial stewardship (AMS) strategies. Methods: Evidence was collected through a narrative literature review using PubMed, WHO, and ECDC sources published between 2010 and 2025. Key terms included “rapid molecular diagnostics,” “sepsis,” “ICU,” “UNYVERO,” “GeneXpert,” “BioFire,” and “carbapenem resistance.” Studies were selected based on clinical relevance to rapid diagnostics and MDR pathogens; no PRISMA-based systematic methodology was applied. Results: Diagnostic performance varies by platform and clinical syndrome. UNYVERO Hospitalized Pneumonia panel demonstrates a sensitivity range of 88.8–91.4% and specificity of 94.9–99.5% in respiratory infections, with a turnaround time of approximately 4–5 h. The GeneXpert Carba-R assay identifies major carbapenemases within 45–60 min with reported sensitivity 96–100% and specificity of 93–99%. BioFire^® Pneumonia and Blood Culture Identification panels similarly provide rapid syndromic results within 1 h, enabling earlier optimization of antimicrobial therapy. Local ICU data from Western Romania identified a substantial burden of carbapenem-resistant Acinetobacter baumannii, underscoring the need for rapid resistance detection to guide therapy. Conclusions: Rapid molecular diagnostics, when integrated with prevention bundles and AMS programs, facilitate earlier targeted therapy, support responsible antimicrobial use, and improve clinical decision-making in MDR infections. Their value is amplified in settings with high resistance prevalence. Wider implementation, combined with surveillance and access to novel antimicrobials, is essential to improve outcomes in critically ill patients. Full article

Background and Objectives: Catheter-associated urinary tract infections (CAUTIs) caused by extended-spectrum beta-lactamase (ESBL)-producing and carbapenem-resistant Enterobacterales (CRE) significantly contribute to global rates of UTI. This study aimed to compare the prevalence and trends of ESBL-producing Enterobacterales and CRE in patients with CAUTIs and non-CAUTIs. Materials and Methods: A retrospective review of 4262 UTI-positive urine cultures was conducted at King Fahad Hospital of the University, Al Khobar, Saudi Arabia (January 2022–November 2023). Demographic, clinical, and microbiological data were obtained from hospital records. Antimicrobial susceptibility was tested using the Vitek^® System; ESBL and CRE were identified using Ezy MIC™ strips and Xpert^® Carba-R assay, respectively. Results: ESBL-producing Enterobacterales accounted for 11.3% of cases; CRE comprised 1.8%. ESBL was significantly more prevalent in non-catheterized patients and those in emergency care. CRE was significantly associated with catheterized patients and inpatient settings. Escherichia coli and Klebsiella pneumoniae were the predominant ESBL-producing and CRE isolates, respectively. bla-OXA-48 was the most frequently detected carbapenemase gene (66.7%). ESBL was prevalent in younger, non-catheterized females, suggesting increasing community transmission. Conversely, CRE were primarily observed in older, catheterized inpatients, emphasizing the role of invasive devices in resistance spread. Conclusions: These findings highlight the importance of targeted infection control and early catheter removal to mitigate resistance trends. Full article

Background/Objectives: The rapid identification of carbapenemase genes directly from positive blood culture (BC) samples shortens the time needed to initiate optimal antimicrobial therapy for Carbapenemase-Producing Enterobacterales (CPE) infections. Several commercial automated PCR systems are available for detecting CPE resistance genes but are expensive. The Xpert^® Carba-R assay (Cepheid GeneXpert System) has high sensitivity and specificity for the detection of carbapenamase genes from bacterial colonies or rectal swabs, with an affordable price. This assay was not used for positive BC testing of CPE resistance genes. Whole-Genome Sequencing (WGS) for resistance genes can be used as the gold standard at a research level. In this study, we evaluated the performance of the Xpert^® Carba-R assay for the early detection of carbapenamase genes directly from positive BCs, using WGS as the gold standard. Methods: A prospective observational study was conducted at Children’s Cancer Hospital-Egypt (CCHE-57357). All positive BCs underwent direct gram staining and conventional cultures. A total of 590 positive BCs containing Gram-negative rods (GNRs) were identified. The Xpert^® Carba-R assay was used to detect carbapenemase genes directly from the positive BC bottle compared with WGS results. Results: Among the 590 GNR specimens, 178 were found to carry carbapenemase genes using the Xpert^® Carba-R assay, with results obtained in approximately one hour. The main genotypes detected were bla_NDM, bla_OXA-48-like, and dual bla_NDM/bla_OXA-48-like at 27%, 29%, and 33%, respectively. The agreement between Xpert^® Carba-R assay and WGS results was almost perfect for the genotype resistance pattern of isolates and individual gene detection. Conclusions: The use of the Xpert^® Carba-R assay directly from BC bottles was an easy-to-use, time-saving, affordable tool with high accuracy in identifying carbapenemase genes and, thus, shortens the time needed to initiate optimal antimicrobial therapy for CPE infections. Full article

Background: Tuberculosis (TB) remains a leading cause of death among people living with HIV (PLWH), yet diagnostic methods vary in accuracy, accessibility, and implementation. Understanding how diagnostic modality influences TB detection is essential to optimizing co-infection management. Methods: We conducted a retrospective analysis of institutional data from Bamrasnaradura Infectious Diseases Institute (BIDI), Thailand, covering 2016–2023. TB detection rates were assessed across five diagnostic methods—chest radiography (CXR), smear microscopy, acid-fast bacilli (AFB) staining, culture, and GeneXpert MTB/RIF—relative to annual HIV-related visit volumes. Results: Among 56,599 HIV-related visits, TB detection rates varied substantially by diagnostic method. CXR was the most commonly used tool, detecting TB in up to 99 cases out of 6964 visits (1.42%) in 2016, though declining to 23 cases out of 6947 visits (0.33%) in 2023. GeneXpert was employed more consistently, yielding between 7 cases out of 7577 visits (0.09%) and 13 cases out of 6593 visits (0.20%) annually. Smear microscopy and AFB staining declined markedly, falling below 0.22% after 2020. These patterns reflect a gradual transition toward molecular diagnostics, which offer improved accuracy but remain underutilized in lower-tier settings. To address these gaps, we incorporated trend analyses confirming significant temporal shifts and propose a tiered TB screening framework tailored to resource availability across healthcare levels. Conclusions: TB detection among PLWH is strongly influenced by the diagnostic method used. Unlike HIV diagnosis—which is definitive and standardized—TB diagnosis remains fragmented and resource-dependent. Context-sensitive screening protocols are urgently needed to improve TB case detection and management, particularly in lower-level HIV care facilities. Full article

Chronic myeloid leukemia (CML) is a clonal myeloproliferative disorder caused by the BCR::ABL1 fusion gene, resulting from a reciprocal translocation between chromosomes 22 and 9. Quantification of BCR::ABL1 transcript levels in peripheral blood by RT-qPCR represents the gold standard for molecular response (MR) monitoring, providing essential clinical information on treatment efficacy. Xpert^® BCR-ABL Ultra is a fully automated in vitro diagnostic test that quantitatively detects e13a2 and e14a2 BCR::ABL1 transcripts using a single-use cartridge that integrates RNA extraction, cDNA synthesis, nested real-time PCR, and signal detection within a rapid, closed, and user-friendly system. In this study, we evaluated Xpert^® BCR-ABL Ultra as an alternative to validated systems currently used by four highly specialized Italian laboratories affiliated with the Italian national laboratory network for CML. A total of 129 peripheral blood samples from CML patients at various disease stages, along with two external quality control materials, were analyzed. We assessed the test’s repeatability, specificity, and stability. Concordance of BCR::ABL1%IS values generated by the different methods was evaluated using EUTOS criteria and Bland–Altman analysis. Finally, MR value concordance was analyzed based on European LeukemiaNet recommendations or calculated using the formula 2 − log₁₀(BCR::ABL1%IS). Xpert^® BCR-ABL Ultra demonstrated high repeatability and stability. The BCR::ABL1%IS values obtained with this assay showed strong concordance with those generated by local reference methods, and MR classifications were consistent across platforms. These findings confirm the robustness, accuracy, and efficiency of the Xpert^® BCR-ABL Ultra assay, supporting its use as a reliable alternative to currently validated systems for the routine clinical monitoring of CML patients. Full article

Background: In 2023, tuberculosis (TB) caused 1.25 million deaths globally, remaining a leading infectious killer. Central Asia and Southern Caucasus face high TB burdens, particularly Mongolia. This review synthesizes TB prevalence data and diagnostic capabilities in these regions to support public health strategies. Methods: This systematic review aimed to synthesize current data on TB prevalence in Central Asia, Southern Caucasus, and Mongolia to support public health strategies and research priorities. A comprehensive search of PubMed and Google Scholar was conducted for English-language articles published up to 2023. Studies were assessed using a modified Newcastle–Ottawa Scale. Nine studies met the inclusion criteria, covering Kazakhstan, Kyrgyzstan, Uzbekistan, Tajikistan, Turkmenistan, Mongolia, Georgia, Armenia, and Azerbaijan. Results: TB incidence ranged from 67 per 100,000 in Kazakhstan to 190 per 100,000 in Kyrgyzstan, with the highest prevalence of 68.5% in Mongolia. TB affected men more frequently (65.3%), and the key risk factors included HIV (30.5%), comorbidities, and undernutrition. Diagnostic performance varied significantly (microscopy sensitivity, 45–65%; GeneXpert MTB/RIF, 89–96% sensitivity and 98% specificity for rifampicin resistance). Diagnostic turnaround times ranged from hours (molecular) to weeks (conventional). Only 58% of TB facilities had GeneXpert technology, with urban–rural disparities in diagnostic access. Drug-resistant TB imposed a significant economic burden, with treatment costs ranging from USD 106 to USD 3125. Conclusions: Strengthening surveillance, improving data collection, and conducting longitudinal studies are essential for designing effective TB control strategies in these regions. Significant diagnostic gaps persist across these regions, especially with regard to drug-resistant strains. Point-of-care molecular diagnostics, improved algorithms, and expanded laboratory training show promise. Future research should focus on rapid biomarker-based diagnostics, field-deployable technologies for settings with limited resources, and AI integration to enhance diagnostic accuracy and efficiency. Full article

Background/Objectives: The concurrent circulation of SARS-CoV-2 with influenza A and B viruses and respiratory syncytial virus (RSV) represents a new diagnostic challenge in the post-COVID-19 area, especially considering that these infections have overlapping clinical presentations but different approaches to treatment and management. Multiplexed molecular testing on point-of-care platforms that focus on the simultaneous detection of multiple respiratory viruses in a single tube constitutes a useful approach for diagnosis of respiratory infections in decentralized clinical settings. This study evaluated the analytical performances of the VitaSIRO solo™ SARS-CoV-2/Flu/RSV Assay performed on the VitaSIRO solo™ Instrument (Credo Diagnostics Biomedical Pte. Ltd., Singapore, Republic of Singapore). Methods: With a view to accreditation, the criteria of the 2022-revised EN ISO 15189:2022 norma were applied for the retrospective on-site verification of method using anonymized respiratory specimens collected during the last 2024–2025 autumn–winter season in France. Results: Usability and satisfaction were comparable to current reference point-of-care platforms, such as the Cepheid GeneXpert^® Xpress System (Cepheid Diagnostics, Sunnyvale, CA, USA). Repeatability and reproducibility (2.34–4.49% and 2.78–5.71%, respectively) demonstrated a high level of precision. The platform exhibited a low invalid rate (2.9%), with most resolving on retesting. Analytical performance on 301 clinical samples showed high overall sensitivities: 94.8% for SARS-CoV-2 (Ct ≤ 33), 95.8% for influenza A and B viruses, 95.2% for RSV, and 95.4% for all viruses. Specificities were consistently high (99.2–100.0%). False negatives (2.6%) were predominantly associated with high Ct values. Agreement with the comparator reference NeuMoDx™ Flu A-B/RSV/SARS-CoV-2 Vantage Assay (Qiagen GmbH, Hilden, Germany) was almost perfect (Cohen’s κ 0.939–0.974), and a total of 91.1%, 94.8%, and 100.0% of Ct values were within the 95% limits of agreement for the detection of SARS-CoV-2, influenza A and B viruses, and RSV, respectively, by Bland–Altman analyses. Passing–Bablok regression analyses demonstrated good Ct values correlation between VitaSIRO solo™ and NeuMoDx™ assays, with a slight, non-significant, positive bias for the VitaSIRO solo™ assay (mean absolute bias +0.509 to +0.898). Conclusions: These findings support VitaSIRO solo™ Instrument as a user-friendly and reliable point-of-care platform for the rapid detection and differentiation of SARS-CoV-2, influenza A and B viruses, and RSV responding to the EN ISO 15189:2022 criteria for accreditation to be implemented in hospital or decentralized settings. Full article

Background: Acute respiratory distress syndrome (ARDS) secondary to tuberculosis (TB) is rare and associated with high mortality. Management is further complicated by comorbidities and ICU-related complications. Methods: We report a 43-year-old woman with post-polio sequelae and uncontrolled diabetes who developed ARDS due to pulmonary TB, complicated by recurrent nosocomial infections and gastrointestinal bleeding. Early bronchoscopy and GeneXpert MTB/RIF PCR were performed on ICU Day 2, enabling anti-TB therapy initiation by ICU Day 3. The patient received lung-protective ventilation, prone positioning, tailored antibiotics, and multidisciplinary care. Results: The patient’s clinical course was complicated by two episodes of ventilator-associated pneumonia and gastrointestinal bleeding, but with individualized management, she achieved ventilator weaning and functional recovery. Conclusions: Early TB recognition in ARDS is crucial. Multidisciplinary ICU management, including prudent steroid use, improves outcomes. Full article

Background/Objectives: High rates of sexually transmitted infections (STIs) increase HIV transmission risk among adolescent girls and young women (AGYW) in South Africa. AGYW prefer discreet self-testing options for HIV and pregnancy; however, other STI self-testing options are currently unavailable in this region. Methods: Seven Chlamydia trachomatis (CT), Neisseria gonorrhea (NG) and Trichomonas vaginalis (TV) assays were validated for AGYW self-test use (using self-collected vaginal samples) in a cross-sectional study (PROVE). Paired GeneXpert^® NG/CT (Cepheid_®, Sunnyvale, CA, USA) and OSOM^® Trichomonas test (Sekisui Diagnostics, Burlington, MA, USA) results from nurse-collected samples served as reference results to calculate sensitivity, specificity, positive predictive values (PPV), and negative predictive values (NPV). One test, the polymerase chain reaction (PCR)-based Visby Medical™ Sexual Health Test device (Visby Medical™, San Jose, CA, USA), was validated for accuracy of positive test results using self-collected samples and home-based testing in a longitudinal follow-up study enrolling AGYW aged 16–18 years. Paired GeneXpert^® NG/CT and TV results from nurse-collected vaginal samples served as reference tests. Results: In PROVE, 146 AGYW contributed 558 paired samples. The Visby Medical™ Sexual Health Test exhibited moderate to high sensitivity (66.7–100%), specificity (80–100%), NPV (66.7–100%), and PPV (66.7–100%) for NG, CT, and TV. The remaining tests’ performances were markedly lower. In the longitudinal study, 28 AGYW contributed 84 paired samples, and the Visby Medical™ Sexual Health Test demonstrated 100% accuracy of positive results for CT, NG, and TV. Conclusions: The Visby Medical™ Sexual Health Test demonstrated high reliability as a potential option for AGYW to discreetly self-test for multiple STIs concurrently. Testing of its acceptability, utility, and feasibility in a larger sample of AGYW is in progress. Full article

Tuberculosis (TB), primarily caused by Mycobacterium tuberculosis complex (MTBC), remains a global health challenge and can lead to severe pulmonary and extrapulmonary complications. Multidrug-resistant TB (MDR-TB) poses additional challenges, requiring advanced diagnostic and treatment strategies. This study evaluates the BD MAX MDR-TB molecular test for a rapid diagnosis of MDR-TB, detecting resistance to rifampicin (RIF) and isoniazid (INH). The BD MAX MDR-TB test, utilizing real-time PCR, was used to analyze specimens collected from TB-suspected patients, identifying MTB DNA and mutations associated with rifampicin and isoniazid resistance. Results were compared with traditional drug susceptibility testing, and 79 out of 638 samples tested were positive for MTB DNA, with 65 showing a sufficient amount of genetic material for resistance gene identification. The BD MAX test showed a 100% correlation with phenotypic rifampicin resistance, though discrepancies were noted for isoniazid resistance, with a 93% concordance. The BD MAX MDR-TB test is an effective tool for a rapid diagnosis of MDR-TB, especially for rifampicin resistance. However, it may not detect certain mutations related to isoniazid resistance. Complementary tests like Xpert MTB/XDR or whole-genome sequencing could improve diagnostic accuracy and support more effective TB control strategies. Full article

Background: Bloodstream infections continue to pose a serious global health threat due to their high morbidity and mortality, further worsened by rising antimicrobial resistance and delays in starting targeted therapy. This study assesses the accuracy and timeliness of therapeutic recommendations produced by an artificial intelligence (AI)-driven and machine-learning (ML) clinical decision support system (CDSS), comparing results based on molecular diagnostics alone with those that combine molecular and phenotypic data (standard cultures). Methods: In a prospective cross-sectional study conducted in Lima, Peru, 117 blood cultures were analyzed using FilmArray/GeneXpert for molecular identification and MALDI-TOF/VITEK 2.0 for phenotypic profiling. The AI/ML-based CDSS provided treatment recommendations in two formats, which were assessed for concordance and turnaround time. Results: Therapeutic recommendations showed 80.3% consistency between data types, with 86.3% concordance in pathogen and resistance detection. Notably, molecular-only recommendations were delivered 29 h earlier than those incorporating phenotypic data. Escherichia coli was the most frequently isolated pathogen, with a 95% concordance in suggested therapy. A substantial agreement was observed in treatment consistency (Kappa = 0.80). Conclusions: These findings highlight the potential of using AI-powered CDSS in conjunction with molecular diagnostics to accelerate clinical decision-making in bacteremia, supporting more timely interventions and improved antimicrobial stewardship. Further research is warranted to assess scalability and impact across diverse clinical settings. Full article